Homo- and Heteroassociations Drive Activation of ErbB3

Tímea Váradi, Magdalena Schneider, Eva Sevcsik, Dominik Kiesenhofer, Florian Baumgart, Gyula Batta, Tamás Kovács, René Platzer, Johannes B. Huppa, János Szöllősi, Gerhard J. Schütz, Mario Brameshuber, Peter Nagy

Research output: Contribution to journalArticle

Abstract

Dimerization or the formation of higher-order oligomers is required for the activation of ErbB receptor tyrosine kinases. The heregulin (HRG) receptor, ErbB3, must heterodimerize with other members of the family, preferentially ErbB2, to form a functional signal transducing complex. Here, we applied single molecule imaging capable of detecting long-lived and mobile associations to measure their stoichiometry and mobility and analyzed data from experiments globally, taking the different lateral mobility of monomeric and dimeric molecular species into account. Although ErbB3 was largely monomeric in the absence of stimulation and ErbB2 co-expression, a small fraction was present as constitutive homodimers exhibiting a ∼40% lower mobility than monomers. HRG stimulation increased the homodimeric fraction of ErbB3 significantly and reduced the mobility of homodimers fourfold compared to constitutive homodimers. Expression of ErbB2 elevated the homodimeric fraction of ErbB3 even in unstimulated cells and induced a ∼2-fold reduction in the lateral mobility of ErbB3 homodimers. The mobility of ErbB2 was significantly lower than that of ErbB3, and HRG induced a less pronounced decrease in the diffusion coefficient of all ErbB2 molecules and ErbB3/ErbB2 heterodimers than in the mobility of ErbB3. The slower diffusion of ErbB2 compared to ErbB3 was abolished by depolymerizing actin filaments, whereas ErbB2 expression induced a substantial rearrangement of microfilaments, implying a bidirectional interaction between ErbB2 and actin. HRG stimulation of cells co-expressing ErbB3 and ErbB2 led to the formation of ErbB3 homodimers and ErbB3/ErbB2 heterodimers in a competitive fashion. Although pertuzumab, an antibody binding to the dimerization arm of ErbB2, completely abolished the formation of constitutive and HRG-induced ErbB3/ErbB2 heterodimers, it only slightly blocked ErbB3 homodimerization. The results imply that a dynamic equilibrium exists between constitutive and ligand-induced homo- and heterodimers capable of shaping transmembrane signaling.

Original languageEnglish
Pages (from-to)1935-1947
Number of pages13
JournalBiophysical journal
Volume117
Issue number10
DOIs
Publication statusPublished - Nov 19 2019

    Fingerprint

ASJC Scopus subject areas

  • Biophysics

Cite this

Váradi, T., Schneider, M., Sevcsik, E., Kiesenhofer, D., Baumgart, F., Batta, G., Kovács, T., Platzer, R., Huppa, J. B., Szöllősi, J., Schütz, G. J., Brameshuber, M., & Nagy, P. (2019). Homo- and Heteroassociations Drive Activation of ErbB3. Biophysical journal, 117(10), 1935-1947. https://doi.org/10.1016/j.bpj.2019.10.001